Method and device for investigating substance libraries

ABSTRACT

The aim of the invention is to investigate the bonding of substances to target molecules. This is achieved by means of a densely packed device wherein various target molecules are bonded in a large number of sample areas. Cross-contamination and evaporation need to be minimized. The active surface of the sample areas have to be maximized. The inventive solution resides in the use of carrier plates, containing densely packed capillary structures and having a very large inner surface despite small outer dimensions. 1000 times more molecules can be bonded than on a flat outer surface of a comparable size. Cross contamination is avoided by the lack of cross links between the capillaries. Evaporation is minimized by a small outer surface. After the inner surface of the capillaries has been silanized, peptides and peptidomimetics are synthesized in a locally targeted manner. The molecular interactions of components of the substance library with active substances in a solution or a suspension are investigated by means of a local resolution optical detection method. Handling, especially cleaning and covering with substances, is carried out in a simple manner by rinsing liquids through the capillary plate.

BACKGROUND ART

[0001] In contrast to solid-phase immobilised miniaturised nucleic acidlibraries, about which there are a large number of publications intechnical journals and patents (S. P. A. Fodor et al. (1991),Light-directed, spatially addressable parallel chemical synthesis.science 251.767-773; E. M. Southern et al. (1992), Analyzing andcomparing nucleic acid sequences by hybridization to arrays ofoligonucleotides: evaluation using experimental models. Genomics13:1008-1017; G. McGall. et al. (1996), Light-directed synthesis ofhigh-density oligonucleotide arrays using semiconductor photoresists,Proc. Natl. Acad. Sci. USA 26:13555-13560, M. Chee et al. (1996),Accessing genetic information with high density DNA arrays. Science274:610-614; 5. Singh-Gasson et al. (1999), Maskiess fabrication oflight-directed oligonucleotide microarrays using a digital micromirrorarray. Nat. Biotechnol. 17:974-978; S. P. A. Fodor et al. (1995), Arraysof materials attached to a substrate. U.S. Pat. No. 5,744,305; S. P. A.Fodor et at. (1995), Very large scale immobilised polymer synthesis.U.S. Pat. No. 5,424,186; G. H. McGall et al. (1995),Spatially-addressable immobilization of oligonucleotides and otherbiological polymers on surfaces. U.S. Pat. No. 5,412,087, A. S.Heuermann (1999), Method and device for photolithographic production ofDNA, PNA and protein chips. WO 9960156A2, G. H. McGall und N. Q. Nam(2000), Synthesis of oligonucleotide arrays usingphotocleavableprotecting groups. U.S. Pat. No. 6,022,963) solid-phaseimmobilised greatly miniaturised peptide or peptidomimetic librarieshave hitherto been described only in a small number of works (S. P. A.Fodor et al. (1991), light-directed, spatially addressable parallelchemical synthesis. Science 25 1:767-773; C. P. Holmes et al. (1995),The use of light-directed combinatorial peptide synthesis in epitopemapping. Biopolymers 37:199-211; M. C. Pirrung et al. (1992), Largescale photolithographic solid phase synthesis of polypeptides andreceptor binding thereof U.S. Pat. No. 5,143,854, M. C. Pirrung et al.(1995), Large scale photolithographic solid phase synthesis of an arrayof polymers. U.S. Pat. No. 5,405,783).

[0002] One of the main reasons why miniaturised peptide libraries,so-called ‘peptide chips’, have hitherto not yet achieved widespreaddissemination, is the relatively high level of expenditure involved inthe production of such arrangements if the conventionalphotolithographic synthesis methods used in relation to ‘DNA chips’ onplanar supports are used. In the case of nucleic acids, those methodsmake use of the fact that there are only four different naturallyoccurring nucleotides (deoxyadenosine, deoxycytidine, deoxyguanosine andthymidine) for producing the deoxyribonucleic acid oligomers, that thecoupling times when forming the oligonucleotides are relatively short(less than 30 minutes) and the yields of the individual coupling stepsare very good (more than 99%). All three criteria have a crucialinfluence on the production time for such chips and thus the economy ofthe entire process.

[0003] In the photolithographic synthesis of oligonucleotides, firstly asupport which is completely protected with a photolabile protectivegroup is cleared of protection in positionally directed fashion byirradiation with light at the locations at which for example a thymidinephosphate is to be applied, and then the entire support is incubatedwith thymidinephosphoamidite which is photolabily protected at the 5′-OHand suitable coupling reagents. Coupling is thus effected only at thedesired locations and the entire procedure has to be repeated for theother three nucleotides before the first dinucleotide can besynthesised. In the case of a chip which is occupied with 20-meroligonucleotides 80 coupling, washing and protection-removal cycles aretherefore required.

[0004] In the case of a peptide however the number of components whichcan be used for coupling is markedly higher than in the case of thenucleic acids. There are 20 proteinogenic amino acids, some naturallyoccurring non-proteinogenic amino acids (for example ornithine), thesame number of corresponding D-amino acids and a continuously risingnumber of artificial amino acids such as for example cyclohexylalanine,aminoisobutyric acid, Norvaline, etc, also each in D-and L-form. Insummary it can be assumed that at the present time about 100 differentamino acids are available for the chemical synthethis of peptides andpeptidomimetics. If—considered conservatively—only half of thosereagents are used for the synthethis of a substance library, that gives1000 coupling and protection-removal cycles in the synthesis of a 20-merpeptide or peptidomimetic library. In addition solid-phase peptidesynthesis generally affords coupling yields of 85-90% with reactiontimes of about 30 minutes so that usually at least one repetition of thecoupling step with fresh reagents is necessary to achieve the requiredsynthesis yields. This means that extremely long synthesis times ofweeks up to several months have to be calculated into the procedure forthe production of a peptide or peptidomimetic library immobilised on atwo-dimensional support if operation is implemented usingphotolithographic methods. Consequently hitherto only photolithographicsyntheses of short peptides of low sequence variability have beendescribed (S P A Fodor et al (1991) Light-directed, spatiallyaddressable parallel chemical synthesis, Science 251;767-773, C P Holmeset al (1995), The use of light directed combinatorial peptide synthesisin epitope mapping. biopolymers 37:199-211; M C Pirrung et al (1992),Large scale photolithographic solid phase synthesis of polypeptides andreceptor binding thereof U.S. Pat. No. 5,143,854).

[0005] An alternative method which considerably reduces the number ofworking steps is based on the simultaneous protection removal of allsupport-bonded reaction partners followed by parallel or sequentialpositionally directed application of the different amino acid reactionmixtures into defined sample areas. In that way it is also possible tosynthesise libraries from longer and complex peptides or peptidomimeticsin an acceptable time frame. The main problem in this respect however isthe cross-contamination which is to be expected of the reactants if thesample areas are close together. Dense packing of the sample areas inturn is desirable in order sufficiently to miniaturise the substancelibrary.

[0006] One possible way of resolving the problem involves increasing thesurface tension in the regions between the sample areas of the planarsupport so that the reaction mixtures remain in the form of smalldroplets in the region of the sample areas. To achieve that aim thesupport surface must be fluoroalkylated between the sample areas, asdescribed in T M Brennan (1995) Method and apparatus for conducting anarray of chemical reactions on a support surface, U.S. Pat. No.5,474,796; T M Brennan (1997), Method and apparatus for conducting anarray of chemical reactions on a support surface, EP 703 825B1 and T MBrennan (1999), Method and apparatus for conducting an array of chemicalreactions on a support surface, U.S. Pat. No. 5,985,551. The methodhowever suffers from the disadvantage that without a protectiveenclosure the drops are severely subjected to evaporation, which canrepresent a major problem particularly when dealing with very smallsample volumes.

[0007] An alternative way of resolving the cross-contamination problemlies in the use of a porous membrane which immediately sucks up theamino acid reaction mixture at the location of application (R Frank(1992), Spot synthesis: an easy technique for the positionallyaddressable, parallel chemical synthesis on a membrane support, Tetrahedron 48:92 17-9232; R Frank and S Guler (1992), Verfahren zurschnellen Synthese von trägergebundenen oder freien Peptiden oderOligonukleotiden, damit hergestelltes Flachmaterial, Verwendung sowieVorrichtung zur Durchfuhrung des Verfahrens, [Method of fast synthesisof support-bonded or free peptides or oligonucleotides, flat materialproduced therewith, use and apparatus for carrying out the method], WO92/04366; J Schneider-Mergener (1994), Verfahren zur Synthese undSelektionierung von Sequenzen aus kovalent verbundenen Bausteinen,[Method of synthesising and selecting sequences of covalently joinedcomponents], WO 94/20521). The unordered capillaries in the porousmembrane, with increasing miniaturisation of the sample areas, resulthowever in cross-contamination and the relatively large surface area ofthe membrane also results in this method in evaporation of the solventand thus under some circumstances in an adverse influence on thecoupling reaction. A further problem which occurs when using thecellulose membranes usually employed here is in part very greatheterogeneity of the synthesised product. In a mass-spectroscopicinvestigation of the entire material detached from a sample area, bothamino-terminally and also carboxy-terminally shortened peptides arefound, which occur due to the fact that the synthesis breaks down tooearly for steric reasons within the membrane and chain re-starts canstill take place by virtue of esterification of amino acids directly atthe cellulose substrate even in later synthesis cycles. In particularpeptides which have been carboxy-terminally shortened by 1-4 amino acidsare demonstrated (D Goehmann and A Frey, unpublished results). Completeblocking of the hydroxyl groups of the cellulose by acetylation orsimilar measures for preventing chain re-starts is prohibited as in thatway the support achieves a higher degree of solubility in the solventsusually employed for solid-phase peptide synthesis and loses aconsiderable amount of mechanical stability (D Goehmann and A Frey,unpublished results). A further disadvantage in the use of membraneslies in their optical properties. In particular cellulose membranes havestrong inherent fluorescence in the emission range of many commerciallyavailable fluorophores so that the sensitivity of identification ofactive substances which bind to the target molecules is greatly reduced(J Helfmann, unpublished results).

[0008] A further approach for resolving the problem ofcross-contamination in charging a planar support with a substancelibrary involves using a microreactor system in which the planar supportrepresents a vessel wall for a plurality of reaction cavities and at thesame time the support medium for the target molecules. The reactioncavities which are enclosed on all sides in that way are specificallysupplied with the desired reagents and washing solutions for example bychanging the positioning of the microreactor block or by means ofmicropassages, electro-osmotic pumps and microvalves in the reactorblock. Waste products are removed in a similar manner (J L Winkler et al(1995), Very large scale immobilised polymer synthesis using 10mechanically directed flow paths, U.S. Pat. No. 5,384,261; P J Zanzucchiet al (1997), Method of synthesis of a plurality of compounds inparallel using a partitioned solid support, U.S. Pat. No. 5,643,738; P JZanzucchi et al (1998), Partitioned microelectronic device array, U.S.Pat. No. 5,755,942; S C Cherukuri et al (1999), Method and system forinhibiting cross-contamination in fluids of combinatorial chemistrydevice, U.S. Pat. No. 5,980,704). A crucial disadvantage of sucharrangements however is their susceptibility to particulate impuritiesin the reaction solutions, in particular if the passages are only a fewmicrometers in diameter and the direction of flow of the reagents andwaste products in the microreactor block is changed a plurality oftimes. The risk of blockage of the microreactor system is very great inparticular in the case of coupling reactions with carbodiimides as theorganic urea derivatives which are produced in this case have a strongtendency to crystallisation. In the worst case then the completemicroreactor block has to be replaced.

[0009] The existing methods and methods described in the literature orpatents all have serious disadvantages. Either there is not a largesurface area and thus not a high level of detection sensitivity withsmall external dimensions, or the synthesis times are overallexcessively long. Optical detection is severely interfered with due toinherent fluorescence and chain breakages result in a non-homogenoussample. When dense packing of the substance library is involvedcross-contamination between various sample areas is to be observed. Whendealing with small volumes evaporation influences the results. Handlingis complicated or very sensitive in relation to particles.

SUMMARY OF THE INVENTION

[0010] The detection sensitivity for the interaction (for examplebinding) of active substances in the liquid phase with solid-phasebonded target molecules is substantially determined by the number ofsolid-phase bonded target molecules. In the case of an ideally planarsupport the solid phase available for binding is limited to the externalsurface of the support. An optical detection system (for examplefluorescence detection) is however always capable of additionallydetecting a given volume above and below the external surface.

[0011] The subject-matter of the invention is an arrangement and amethod as defined in the claims. Particular configurations of theinvention are claimed in the appendant claims and set forth in thedescription hereinafter without the invention being restricted to theillustrated embodiments.

[0012] The invention generally concerns an arrangement for receiving andbuilding up a substance library and a method of producing such alibrary. In that respect the term ‘substance library’ is used to denotea library or collection of many different but usually similar substancesin the same class. The library usually serves to search through thesubstances belonging thereto on the basis of some property and to findand possibly select therein suitable members or target substances.Examples of substances which can form a substance library according tothe invention and which are also referred to as target molecules areinter alia DNA sequences, peptides, polypeptides and peptoid substances.

[0013] In particular the invention concerns a structure which cancomprise glass, quartz, ceramic, plastic material, semi-metal or metaland which by virtue of a high density of small capillaries 1 (diameters5-100 μm) has a large internal surface area with a multiple of theexternal surface area (see FIG. 1). The overall thickness of thecapillary plate 4 (thickness up to 10 mm) and thus the total length ofthe capillaries 1 and the surface thereof is detected by a detectionapparatus. As the maximum occupation (number of molecules) of thecapillary plate 4 is determined by the size of the internal surfacearea, and that is larger by a multiple (between 100 and 1000 times) incomparison with the external surface area, the detection sensitivity isincreased to the same degree when using the capillary plate 4, incomparison with a planar sample arrangement.

[0014] Surprisingly it has been found that peptide and peptoid moleculescan be synthesised on a silanised surface of high quality and the numberof chain re-starts can be almost completely eliminated. If in addition aprolonged anchor molecule which serves as a spacer is also coupledbetween the surface and the target molecule, the chain breakages aregreatly reduced and accessibility for the active substances isconsiderably improved.

[0015] To produce a library of target molecules, from which eachindividual species or defined mixtures of given species must besynthesised or bonded to previously defined regions of the capillaryplate 4, firstly the internal surface of the capillary plate 4 must beenabled for covalent bonding of the target molecules. That is effectedby applying an organosilane layer which has functional groups foranchoring peptidic or peptoid target molecules. Preferably aγ-aminopropyltrialkoxysilane is used for that purpose, however otherorganofunctional silanes such as for exampleγ-mercaptopropyltrialkoxysilane are also suitable for that purpose. Whenusing metal or semi-metal capillary plates an oxide layer must beproduced first for binding the silane by means of surface oxidation ofthe capillary plate.

[0016] Then, a respective sample area 3 is applied to thatorganofunctionalised capillary plate 4 by position-dependent applicationof a substance (for example by pipetting). In the simplest case thesubstance can be a reagent which temporarily protects the functionalgroups of the organosilane layer intended for binding. In a preferredembodiment however an anchor molecule is applied in the sample areas 3,with which the target molecules can be pushed between 0.5 and 20 nmbeyond the internal surface of the capillary plate 4 in order in thatway to permit better active substance binding. The anchor reagent usedis for example fluorenylmethoxycarbonyl (FMOC)-protectedα-amino-poly(ethyleneglycol)-ω-propionic acid-N-hydroxybenzotriazoleester.

[0017] Then, all regions of the capillary plate 4 which are notderivatised with protection or anchor reagents are chemically sosaturated that neither synthesis or coupling of target molecules nornon-specific binding of active substances can occur there at a latertime. Chemical saturation of the non-sample areas can be effecteddepending on the respective requirements involved with hydrophilic,hydrophobic or oleophobic groups. Saturation is effected with acetylresidues under standard conditions.

[0018] In the following steps for production according to the inventionof a substance library which is also composed of a plurality of sampleareas each with a respective species of target molecules, the protectivegroup on the support or the anchor molecules is synchronously removed inall sample areas 3 and then coupling or synthesis of the peptidic andpeptoid target molecules is effected in the sample areas 3 provided forsame.

[0019] Knowledge of the substance which is bonded or synthesised at eachlocation in that way permits parallel analysis of the interactions ofvarious target molecules with applied active substances. A prerequisitein that respect is that no mutual contamination occurs in the operationof position-dependently applying or synthesising the target molecules.

[0020] In a development of the concept of the invention, that isachieved by the parallel arrangement of the capillaries 11 which extendperpendicularly to the plate surface and which are only open towards theexternal plate surfaces. The absence, which is achieved in that way, ofany cross-connections between various sample areas 3 successfullyprevents cross-contamination phenomena, even when the sample areas 3involve a high level of density. A sample area 3 includes a plurality of(for example up to 4000) capillaries 1. Substances which are appliedthereto by pipetting are drawn into the capillaries 1, which can also beassisted depending on a respective liquid involved by the production ofa reduced pressure on a capillary plate side. Flow out of thecapillaries 1 by way of a plurality of capillaries 1 into another samplearea 3 is out of the question as the liquid is held in the capillary bycapillary force. With these liquids, solutions are effectively preventedfrom dripping out of the capillaries 1, due to the very low viscosityinvolved, by a plate 12 (FIG. 2) with a hydro- and oleophobic surface,which is positioned to the underside of the capillary plate 4.

[0021] In a further development of the concept of the invention, byvirtue of the capillaries 1 which are open towards both external platesurfaces, simple automation of a solid-phase peptide synthesis procedurewhich is implemented in the capillary plate 4 is also possible. In thatrespect, after the conclusion of a coupling step the reagents and wasteproducts can be sucked out of the capillaries 1 unidirectionally byapplying vacuum to the underside of the capillary plate 4 and thecapillaries 1 are synchronously flushed by applying washing solutions onthe top side of the capillary plate 4. For prolonged incubationprocedures and washing steps, all reagents and washing solutions whichare to be synchronously applied to all regions of the capillary plate 4can also be brought into contact with all capillaries 1 by flooding theentire capillary plate 4 from the underside, and then removed again byapplying reduced pressure. An apparatus for the synchronous applicationof washing and reagent solutions to all regions of the capillary plate 4is shown in FIG. 2.

[0022] After different target molecules have been synthesised or bondedon the target areas 3 in position-dependent fashion in that way, theentire support is exposed to a dissolved or suspended particulate activesubstance in the liquid phase, in which respect it is necessary toensure that, when using suspensions, the particle size is alwaysmarkedly below the inside diameter of the capillaries 1. The redundancywith many capillaries per sample area however means that individualparticles do not represent a risk of blockage for an entire sample area.Incubation of the substance library with active substance solutions orsuspensions and flushing operations before and after same are carriedout in a preferred embodiment involving a simple structure, as is shownin FIG. 3.

[0023] Analysis of active substance binding is effected in a preferredembodiment optically by means of fluorescence markings by binding afluorophore to the active substances. In an alternative embodiment thefluorophores are bonded to the target molecules. For reading-outpurposes, the individual sample volume which is composed of thethickness of the capillary plate and the sample region 3 is completelypenetrated by excitation light and also completely detected by thedetection optics. The plurality of sample locations is detected bytime-sequential rastering, in which case either the capillary plate ismoved two-dimensionally under the stationary optical arrangement or thebeam path is moved over the capillary plate. In accordance with theinvention the use of quartz or glass as the material for the capillaryplate here minimises the influence of

[0024] Detailed Description of the Solution According to the Inventionin a Preferred Embodiment

[0025] The way in which the entire capillary plate is pre-treated is nowdescribed. The sample areas are then produced and the substancessynthesised thereon. An automatic device is described for all proceduresto which the capillary plate is completely exposed, such as for examplewashing. That device is combined with a pipetting robot so that thecapillary plate can be washed and protection removed therefrom in thesame device, and the individual synthesis steps can be implementedtherein. A further device is described for investigating the binding ofan active substance to the substance library, the capillary plate beingfitted into the further device and brought into contact with the activesubstance. Optical detection of the binding effect is carried out at thesame time herein.

[0026] Derivatisation of the Capillary Plate for Receiving the SubstanceLibrary

[0027] The capillary plate is produced by etching from homogeneousglass, in an alternative method from heterogeneous glass. Oxidation ofsilicon and metal surfaces and silanisation of glass, quartz, silicon,ceramic and oxidised silicon or metal surfaces is state of the art andhas already been described many times in the specialist and patentliterature (for example regarding the surface oxidation of a siliconsupport: A W Flounders et al (1997), Patterning of immobilised antibodylayers via photolithography and oxygen plasma exposure, BiosensorsBiolelectronics 12:447-456; for example regarding silanisation: M Lynn(1975), Inorganic support intermediates: covalent coupling of enzymes oninorganic supports, In: ‘Immobilised Enzymes, Antigens, Antibodies andPeptides’, H H Weetall, Hrsg, Marcel Dekker, New York, N.Y., USA, pages1-48; H M Weetall (1976), Covalent coupling methods for inorganicsupport materials, Methods Enzymol 44:134-148).

[0028] In the preferred embodiment for that purpose an untreatedcapillary plate 4 with surface oxide, ceramic or glass coating is freedof adsorbed organic compounds either in an oxidising acid such as forexample chromosulfuric acid or hot concentrated nitric acid, for onehour, washed between five and ten times by sucking through high-puritywater (on a suction filter or a Büchner funnel, glass frit or a rinsingdevice developed especially for the capillary plate 4 (FIG. 2)), anddried at 250° C.

[0029] Alternatively adhering organic compounds can be removed bypyrolysis at 500° C. in an oxygen atmosphere. After cooling thecapillary plate 4 is coated for 18 hours at ambient temperature in asolution of 2% (v/v) γ-aminopropyltriethoxysilane in water/acetone(1:1), washed ten times by sucking through acetone, left to dry andsintered overnight at 120° C.

[0030] Definition of the Sample Areas

[0031] In the preferred embodiment definition of the sample areas 3 is atwo-stage method, wherein firstly portions of a reaction solution withanchor molecules are pipetted by means of a programmable pipetting robotinto given regions of the capillary plate 4 which thus become sampleareas 3. In the second step all reactive groups in the non-sample areasare saturated with acetyl residues.

[0032] For that purpose in each case between about 0.4 and 4000 ml of asolution which contains a double molar excess—with respect to the numberof amino functions in the corresponding region of the derivatisedcapillary plate 4—of α-FMOC-aminopoly(ethyleneglycol)propionicacid-ω-N-hydroxybenzotriazole ester in N-methylpyrolidone, pipetted intothe future sample areas 3 of the capillary plate 4, incubated for forexample four hours at ambient temperature, in each case washed threetimes by sucking through dimethylformamide and absolute ethanol, anddried under clean room conditions in an air flow. The entire couplingand washing cycle can be repeated between one and three times toincrease the coupling yield. Theα-FMOC-aminopoly(ethyleneglycol)propionic acid-ω-N-hydroxybenzotriazoleester is immediately previously produced for example by the reaction ofα-FMOC-aminopoly(ethyleneglycol)propionic acid with2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluoro-phosphate(HBTU) (for example in a ratio of 1:1.2) or in situ by radical esterinterchange of for example α-FMOC-aminopoly(ethyleneglycol)propionicacid-ω-hydroxysuccinimide ester with water-free N-hydroxybenzotriazoleand diisopropylcarbodiimide (in a ratio of 1:0.5:0.5) inN-methylpyrrolidone.

[0033] In the subsequent saturation procedure the entire capillary plate4 is incubated for example in each case three times for one hour in eachcase at ambient temperature in between five and ten times the internalplate volume of a solution of 2% (v/v) acetic anhydride, and 1% (v/v)diisopropylethylamine in dimethylformamide, wherein the plate is movedin the solution every 15 minutes in such a way that the reactionsolution is replaced in the capillaries 1. Thereafter the capillaryplate 4 is washed on a frit or suction filter three times in each casewith dimethylformamide and ethanol and dried in an air flow.

[0034] In the following protection removal procedure the saturatedcapillary plate 4 is incubated three times in each case for 5 minutes atambient temperature in between five and ten internal plate volumes of asolution of 20% (v/v) piperidine in dimethylformamide. Thereafter thecapillary plate 4 is washed and finally dried as described above.

[0035] Production of the Substance Library in the Sample Areas

[0036] Synthesis of the substance library of peptidic or peptoid targetmolecules takes place semi- or fully automatically using the standardFMOC synthesis process for cellulose filter-immobilised peptidelibraries (R Frank (1992), Spot synthesis: an easy technique for thepositionally addressable, parallel chemical synthesis on a membranesupport. Tetra hedron 48:9217-9232), wherein the solutions which containthe activated, amino-terminally and side chain-protected amino acids arepipetted by means of a pipetting robot into the corresponding sampleareas 3 of the capillary plate and there drawn by capillary forces intothe capillaries 1 and incubated for the course of the coupling reaction.In the fully automatic process (see FIG. 2) the capillary plate 4 iscoated with 5 times the internal plate volume of dimethylformamidewithin 30 seconds and the reaction mixtures sucked away, followed by thewashing solution layered thereover, by reduced pressure. Then, eitherthe amino functions which have not reacted are blocked by means ofacetic anhydride or after further washing of the capillary plate 4 indimethylformamide and ethanol the entire coupling cycle is repeatedbetween one and three times before the amino functions which have notreacted are blocked and then the FMOC protection groups are removed inthe sample areas 3 of the entire capillary plate 4. That synthesis cycleis repeated until the peptides are obtained in the desired length. Thenthe side chains of the amino acids have the protection removedtherefrom, the entire capillary plate 4 is washed with dimethylformamideand ethanol, dried under clean room conditions in an air flow, weldedinto a plastic film under protective gas (for example argon or nitrogen)and stored at −70° C. in the presence of drying agent until required forfurther investigation of the binding of active substances to thesubstances on the capillary plate.

[0037] As an alternative thereto it is also possible to apply to thefree amino functions in the sample areas 3 of the capillary plate 4 forexample bromoacetyl residues for coupling thiol-bearing peptides oraldehyde residues for binding proteins. Which bioconjugate chemistry isoptimally suited here depends on the individual requirements of thepeptidic or peptoid target molecules to be coupled and cannot begenerally applicably described at this point.

[0038] Device for Fully Automatic Washing, Saturation and ProtectionRemoval

[0039] Fully automatic washing, saturation and protection removal is thepreferred embodiment in terms of producing the substance library. It ispreferred over a manual method as it saves on working time and reagentsand the entire synthesis procedure is shortened and made morecost-efficient (FIG. 2).

[0040] For mounting purposes the capillary plate 4 is fitted on to theunderside of a synthesis table 6 which is fixedly connected to apipetting robot. In this case the edge regions 5 of the capillary platecome to lie on a soft, solvent-resistant seal 7. Spacers 8 are thenpushed to close to the edge regions 5 around the capillary plate 4 andsecured to prevent them from moving by screwthreaded bolts 9 so that thecapillary plate 4 is stationarily positioned on the sample table.Thereafter the upper part of the synthesis table 10 is fitted on to thescrewthreaded bolts 9 and pulled by means of a nut 11 against thecapillary plate 4 and the spacer 8 or the lower part of the synthesistable 6 so that the seals 7 disposed in the upper and lower parts of thesynthesis table 10 and 6 are compressed and seal off the capillary plate4 towards the side.

[0041] For charging the sample areas 3 the hydrophobically andoleophobically coated piston 12 is pushed against the underside of thecapillary plate 4 so that no reaction solution can issue downwardly fromthe capillary plate 4 during the coupling reaction.

[0042] For the washing operation after termination of the couplingreaction the entire capillary plate 4 is coated with washing solutionfrom the washing solution feed flow means 13, the piston 12 is moveddownwardly and vacuum is applied to remove the solutions sucked throughthe capillary plate 4 at the discharge 14. The piston 12 is then movedback to the underside of the capillary plate 4 again, the capillaryplate 4 is again covered with washing solution and the washing operationis repeated as may be desired (FIG. 2A). After the conclusion of thewashing operation air is sucked through the capillary plate 4 until allsolvent residues are evaporated. The piston is then moved back to thecapillary plate 4 again and a renewed pipetting cycle is carried out,and the plate is saturated or protection removed therefrom. Forsaturation of unreacted amino functions and for protection removal fromamino functions the piston 12 is moved downwardly and the cavity whichoccurs by virtue of retraction of the piston 12 beneath the capillaryplate 14 is filled with the desired reaction solution through thereagent feed flow means 15. When the discharge 14 and the reagent feedflow means 15 are closed the piston is moved slowly up and down for theduration of the reaction so that the reaction solution flows in auniform flow through the capillaries 1 and the reaction can take place(FIG. 2B). The capillary plate 4 is then washed as described above.

[0043] Device for Binding Active Substances to the Substance LibraryImmobilised on the Capillary Plate

[0044] Investigation of the binding of active substances to thesubstance library on the internal surface of the capillary plate 4requires a device with which an active substance solution or suspensioncan be uniformly flushed through all capillaries 1 so that every activesubstance component can come into contact with every target moleculespecies (FIG. 3).

[0045] In the preferred embodiment, for mounting purposes the capillaryplate 4 is laid on the lower part of the incubation table 16. In thatcase the edge regions 5 of the capillary plate come to lie on a softseal 17. Then spacers 18 are laid around the capillary plate 4, theupper part of the incubation table 19 is fitted on and pressure isapplied by a clamp 20 by means of pressure springs 21 to the capillaryplate 4 and the spacer 18 or the lower part of the incubation table 16,so that the seals 17 in the upper and lower parts of the incubationtable 19 and 16 are compressed and seal off the capillary plate 4towards the side. Then an ultrasonic transmitter 22 is pushed into anopening in the spacer 18 until it is in contact with the edge region 5of the capillary plate 4.

[0046] To charge the device the piston 23 is pushed downwardly and thecavity which is formed by retraction of the piston 23 beneath thecapillary plate 4 is filled with the desired active substance solutionor suspension through the filling connection 24. The volume of theactive substance solution or suspension should be at least 1.5 times theinternal volume of the capillary plate 4 plus the volume of the cavity25 above the capillary plate 4.

[0047] For incubation purposes when the discharge 26 and the fillingconnection 24 are closed the piston 23 is moved slowly upwardly so thatthe air can escape from the capillaries 1 into the cavity 25. Air whichis still enclosed in capillaries 1 can be removed by acousticirradiation with the ultrasonic transmitter 22. When the piston 23 ismoved to the capillary plate 4 the cavity 25 is completely filled withactive solution or suspension and both the air and also the superfluousactive substance solution or suspension escape through lateral passages27 into the compensating vessels 28. The solution or suspension in thecapillaries 1 is then mixed with the solution or suspension in thecavity 25 by repeated upward and downward movement of the piston 23 witha short stroke and air bubbles still present in the cavity 25 aredisplaced stepwise into the compensating vessels 28.

[0048] To empty the device the piston 23 is pushed back to such anextent that the discharge 26 is cleared, through which the activesubstance solution or suspension is then removed by applying reducedpressure.

[0049] Washing of the device is effected in a similar manner usingsuitable washing solutions.

[0050] Positionally resolved detection of the active substance reactionwith the target molecules can also take place in the device as thecavity 25 above the capillary plate 4 is delimited upwardly by atransparent window 29 and thus for example binding or cleavage offluorophores in the device can be detected or tracked by means of themeasuring device described with reference to FIG. 4.

[0051] Device for Detecting the Binding of Active Substances to TargetMolecules

[0052] Detection methods for the binding of molecules are described inmany different forms in the literature and in patents, being basedmostly on the use of radioactive or optical markers. Optical methodswhich permit penetration of the branches or webs between the capillariesare used for detecting bindings in the capillary plate. The most variedoptical methods are based on fluorescence markings (for example M VRogers (1997), Light on high-throughput screening: fluorescence-basedassay technologies, Drug discovery Today 2:156-160). Fluorescencemarking is produced by coupling a fluorophore either to the targetmolecules or to the active substances. For reading-out purposes, in thepreferred embodiment the individual sample volume which corresponds tothe sample area 3 over the entire thickness of the capillary plate iscompletely penetrated by excitation light and also completely detectedby the detection optics. In order to prevent cross-talk of adjacentsample areas, only one sample location is illuminated at a moment intime and detection is limited only to one sample location. Themultiplicity of sample locations are detected by time-sequentialrastering. Measurement at a sample location can be effected bothcontinuously and also with repetitive pulses. In the former case,irradiation is effected with a continuously radiating, bandpass-filtered light source or a laser, wherein the narrow-bandexcitation light is adapted to the absorption of the fluorescencemarker. Detection is effected in a narrow-band mode in the range offluorescence emission of the fluorescence marker with a suitablephotodetector. In the case of pulsed fluorescence excitation as shown inFIG. 4, a laser 30 is used for illumination purposes, the pulse durationof which is at least a third of the life of the marking fluorophore. Thecollimated beam of the laser 31 is reflected by a wavelength-selectivebeam splitter 32 in a direction towards the capillary plate 4. The beamis focussed by an objective lens 33 on to a sample area 3 of thecapillary plate 4 in such a way that the radiation passes through theentire sample volume of a sample area 3. The isotropically radiatedfluorescence light 34 of the fluorescence marking is collimated by thesame objective lens 33 and passes through the selective beam splitter 32by virtue of the greater wavelength of the fluorescence. Stray light ofthe excitation laser is greatly suppressed by a narrow-band filter 35,but the fluorescence light is passed at the highest possibletransmission. The fluorescence light is focussed on to the photodetector37 by a further objective lens 36. Time-resolved detection with a risetime of less than a third of the fluorescence life of the fluorophoremeans that the fluorescence signal can particularly advantageously bedetected in a time gate integration procedure with a boxcar integrator38 after decay both of the elastic stray light and also Raman-scatteredlight. The time gate is correspondingly opened approximately afterdouble the laser pulse duration and remains open for approximatelydouble the fluorophore life.

[0053] A trigger signal from the laser 39 supplies the time base for thetime gate. Standardisation of the fluorescence light and thus correctionin respect of fluctuations in the laser pulse energy can be implemented,by way of a second channel of the boxcar integrator 38, by means of asignal from the laser 40, which is proportional to the laser pulseenergy. All data are collected in a computer 41 and finally plotting ofthe fluorescence intensity over the respective sample location can berepresented and association of the detected signals with the targetmolecules can be effected.

DESCRIPTION OF THE FIGURES

[0054]FIG. 1 is a diagrammatic view of a capillary plate 4 in plan andas a cross-section. The individual capillaries 1 are separated from eachother by branch or web regions 2. The sample areas 3 include a pluralityof capillary openings. The edge region of the plate 5 does not have anycapillaries.

[0055]FIG. 2 shows a device for fully automatically washing, saturatingand removing protection from the capillary plate 4 during synthesis ofthe substance library. In this case the capillary plate 4 is fittedbetween the upper part 10 and the lower part 6 of the synthesis tableand fixed in position by means of suitable seals 7 and spacers 8 by wayof a screwthreaded bolt 9 with nut 11. Washing solution feed flow means13 are provided in the upper part 10 of the synthesis table and areagent feed flow means 15 and a discharge 14 are provided in the lowerpart 6. A piston 12 can be moved in the lower part 6 of the synthesistable and can be moved to the underside of the capillary plate 4.

[0056]FIG. 3 shows a device for binding active substances to thesubstance library which is immobilised on the capillary plate. Thecapillary plate 4 is fitted between the upper part 19 and the lower part16 of the incubation table and fixed in position by means of suitableseals 17 and spacers 18 by a clamp 20 with a pressure spring 21. Anultrasonic transmitter 22 is mounted laterally to the capillary plate 4.Disposed in the lower part of the incubation table are a fillingconnection 24 and a discharge 26 for the active substance solutionswhich can be pressed with a piston 23 through the capillary plate 4 intothe upper cavity 25. Lateral passages 27 lead from the cavity 25 into acompensating vessel which serves to catch displaced air and excessactive substance solution. The cavity 25 is closed off at its top by atransparent window 29 so that direct optical detection of reactions inthe device is possible.

[0057]FIG. 4 is a diagrammatic view of a device for detecting thebinding of active substances to the substance library by means ofoptical methods. Fluorophore-marked target molecules on the capillaryplate 4 are excited to fluorescence by a laser beam 31 which is producedby a pulsed diode laser 30 and which is deflected on to the sample areaby a wavelength-selective beam splitter 32 and an objective lens 33. Theradiated fluorescence light 34 passes through the beam splitter 32, aband pass filter 35 and a further objective lens 36 before it impingeson the photodetector 37. For integration of the fluorescence signalwithin a time gate, a boxcar integrator 38 receives a time triggersignal 39 from the laser and a signal 40 proportional to the laserenergy for standardisation of the fluorescence signal. A movable sampletable 42 is so controlled by a computer 41 that the sample areas can besuccessively measured. The standardised fluorescence signal isrepresented over the sample location.

[0058] Legend

[0059]1 capillary

[0060]2 branch region between capillaries

[0061]3 sample area

[0062]4 capillary plate

[0063]5 edge region of the capillary plate

[0064]6 lower part of the synthesis table

[0065]7 solvent-resistant seal

[0066]8 spacer

[0067]9 screwthreaded pin

[0068]10 upper part of the synthesis table

[0069]11 nut

[0070]12 piston

[0071]13 washing solution feed flow means

[0072]14 discharge

[0073]15 reagent feed flow means

[0074]16 lower part of the incubation table

[0075]17 seal

[0076]18 spacer

[0077]19 upper part of the incubation table

[0078]20 clamp

[0079]21 pressure spring

[0080]22 ultrasonic transmitter

[0081]23 piston

[0082]24 filling connection

[0083]25 cavity

[0084]26 discharge

[0085]27 passage

[0086]28 compensating vessel

[0087]29 transparent window

[0088]30 pulsed diode laser

[0089]31 laser beam

[0090]32 beam splitter

[0091]33 objective lens

[0092]34 radiated fluorescence light

[0093]35 band pass filter

[0094]36 objective lens

[0095]37 photodetector

[0096]38 boxcar integrator

[0097]39 trigger pulse from the laser

[0098]40 signal from the laser which is proportional to the laser pulseenergy

[0099]41 computer

[0100]42 movable sample table

[0101] Some preferred configurations of the arrangement according to theinvention and the method according to the invention will also be setforth hereinafter.

[0102] A. Method and device for investigating the molecular interactionof soluble or suspendable active substances with solid-phase bondedpeptidic or peptoid target molecules characterised in that the surfacenecessary therefor is provided within a plate in the form ofcapillaries. The capillaries pass through from one side of the plate tothe opposite side but in so doing are not connected to each other.

[0103] B. A method and device as defined in A which are characterised inthat a plurality of parallel capillaries are arranged in the plate,which draw liquids through capillary force into the capillaries and holdsame therein.

[0104] C. A method and device as defined in the preceding paragraphswhich are characterised in that the capillary plate is made from one ofthe materials glass, quartz, silicon, plastic material, semi-metal,ceramic, metal or a combination of said materials.

[0105] D. A method and device as defined in the preceding paragraphswhich are characterised in that the internal surface is enabled forcovalent bonding of target molecules or synthesis of the targetmolecules is effected.

[0106] E. A method and device as defined in the preceding paragraphswhich are characterised in that the internal surface is covered by anorganosilane layer which has functional groups for anchoring peptidic orpeptoid target molecules.

[0107] F. A method and device as defined in the preceding paragraphswhich are characterised in that γ-aminopropyltrialkoxysilane or otherorganofunctional silanes are used for the silanisation step.

[0108] G. A method and device as defined in the preceding paragraphswhich are characterised in that when using metal or semi-metal capillaryplates prior to the silanisation step an oxide layer is provided forbinding the silane by means of surface oxidation of the capillary plate.

[0109] H. A method and device as defined in the preceding paragraphswhich are characterised in that locally delimited sample areas areapplied in the capillary plate.

[0110] I. A method and device as defined in the preceding paragraphswhich are characterised in that a plurality of sample areas each of adiameter of up to 2000 μm or a number of up to 4000 adjacent capillariesare applied on a capillary plate.

[0111] J. A method and device as defined in the preceding paragraphswhich are characterised in that the capillaries are of a length and thecapillary plate is of a thickness of between 100 and 2000 μm.

[0112] K. A method and device as defined in the preceding paragraphswhich are characterised in that the functional group of the organosilanelayer is temporarily protected by pipetting a substance on to thecapillary plate. The region protected in that way represents a samplearea.

[0113] L. A method and device as defined in the preceding paragraphswhich are characterised in that a protected anchor molecule is producedat each binding location of the sample area by pipetting a reagent on tothe capillary plate, wherein the anchor molecule serves at the same timeas a spacer in relation to the internal surface.

[0114] M. A method and device as defined in the preceding paragraphswhich are characterised in that a protected sample area is applied bypipetting on to the plate fluorenylmethoxycarbonyl (FMOC)-protectedα-aminopoly(ethyleneglycol)-ω-propionic acid-N-hydroxybenzotriazoleester.

[0115] N. A method and device as defined in the preceding paragraphswhich are characterised in that all non-protected regions of thecapillary plate are chemically saturated.

[0116] O. A method and device as defined in the preceding paragraphswhich are characterised in that the protective groups are removed in thesample areas of the entire capillary plate.

[0117] P. A method and device as defined in the preceding paragraphswhich are characterised in that peptides or peptoid target molecules arepositionally dependently sequentially synthesised out of amino acids inthe sample areas of the capillary plate.

[0118] Q. A method and device as defined in the preceding paragraphswhich are characterised in that the complete peptide or peptoid targetmolecules are bonded in positionally dependent manner in the sampleareas of the capillary plate.

[0119] R. A method and device as defined in the preceding paragraphswhich are characterised in that a substance library is built up by thepositionally dependently different sequences of peptide or peptoidtarget molecules.

[0120] S. A method and device as defined in the preceding paragraphswhich are characterised in that a closed and variable volume providedwith a valve exists on at least one side of the capillary plate, intowhich volume liquids with soluble or suspendable active substances andother liquids and gases can be introduced and sucked away.

[0121] T. A method and device as defined in the preceding paragraphswhich are characterised in that gas bubbles can be dissolved by couplingultrasound to the capillary plate.

[0122] U. A method and device as defined in the preceding paragraphswhich are characterised in that the reaction speeds in the capillariesare increased by coupling ultrasound.

[0123] V. A method and device as defined in the preceding paragraphswhich are characterised in that the binding of the active substances tothe target molecules is detected.

[0124] W. A method and device as defined in the preceding paragraphswhich are characterised in that a change in the target molecules can beproduced by the active substances and detected.

[0125] X. A method and device as defined in the preceding paragraphswhich are characterised in that the capillary plate is transparent oroptical and is freely accessible to optical measurement.

[0126] Y. A method and device as defined in the preceding paragraphswhich are characterised in that soluble or suspendable active substancesare fluorescence-marked and introduced into the capillaries.

[0127] Z. A method and device as defined in the preceding paragraphswhich are characterised in that after a given time the non-bonded activesubstances are removed and the bonded active substances are detected onthe basis of fluorescence marking.

[0128] AA. A method and device as defined in the preceding paragraphswhich are characterised in that the solid-phase bonded peptidic orpeptoid target molecules are fluorescence-marked.

[0129] BB. A method and device as defined in the preceding paragraphswhich are characterised in that soluble or suspendable activesubstances, acids or other liquids are flushed into the capillaries,flushed out again after a given time, and the presence of the stillbonded fluorescence-marked target molecules is detected.

[0130] CC. A method and device as defined in the preceding paragraphswhich are characterised in that the fluorescence marking is excited anddetected with light in the UV, in the visible range or near infrared.

[0131] DD. A method and device as defined in the preceding paragraphswhich are characterised in that fluorescence excitation is effected witha pulsed diode laser with a pulse duration of less than 10 ns.

[0132] EE. A method and device as defined in the preceding paragraphswhich are characterised in that detection of fluorescence is effectedfor the suppression of stray light in a time gate after decay of theexcitation pulse.

What is claimed is:
 1. (amended) A device for producing peptidic or peptoid substance libraries on a planar structure that has areas which are laterally separated from each other by physical barriers, for receiving a plurality of individual peptidic or peptoid target molecules or the precursors thereof that define the substance library, the device comprising: a synthesis table for fixing the structure in a defined position during the production method; a pipetting robot which applies the necessary reagents for producing locally delimited sample areas from a respective group of areas and applies the necessary reagents for producing the individual target molecules at the same time or in time-displaced relationship in the respective sample areas of the structure, and a variable-volume cavity with a discharge at an underside of the synthesis table for disposal of applied reagents.
 2. (amended) The device of claim 1, wherein the structure comprises a capillary plate with a plurality of capillaries which serve to receive the individual peptidic or peptoid target molecules of the substance library or the precursors thereof.
 3. (amended) The device of claim 2, wherein at least an internal surface of the capillaries is coated with an organosilane layer which has functional groups suitable for covalent bonding of the target molecules of the substance library.
 4. (amended) The device of claim 3, wherein the structure has regions outside the sample areas with functional groups that are chemically deactivated.
 5. (amended) The device of claim 4, wherein the planar structure is transparent or opaque and can be used for optical measurement.
 6. (amended) A method of building up peptidic or peptoid substance libraries defined by a plurality of individual peptidic or peptoid target molecules or the precursors thereof on a planar structure having areas which are separated laterally from each other by physical parameters, the method comprising the following steps: a) fixing the structure in a defined position on a synthesis table during the production method; b) covering at least the laterally-separated areas of the structure with an organosilane layer which has functional groups for anchoring peptidic or peptoid target molecules; c) defining locally delimited sample areas comprising a group of adjacent areas by selectively applying a reagent for protecting the functional groups or a protected anchor molecule for covalent bonding to the functional groups by a pipetting robot to the areas of the sample areas; d) chemically deactivating the regions of the structure, which are outside the sample areas, with functional groups, by the application of a deactivation reagent; e) deprotecting all protected functional groups or anchor molecules; f) applying the amino-terminally and side chain-protected, reagents required for production of the individual target molecules to the sample areas of the structure selectively with the pipetting robot and covalently bonded to the de-protected functional groups or anchor molecules; g) removing excess reagents by way of a variable-volume cavity with a discharge at an underside of the synthesis table for disposal thereof, and h) repeating steps e) through g) as required.
 7. (amended) The method of claim 6, wherein the structure is a capillary plate with a plurality of capillaries which serve to receive individual peptidic or peptoid target molecules of the substance library or precursors thereof, comprising the following method steps: a) fixing the capillary plate in a defined position on a synthesis table during the production method; b) covering at least the internal surface of the capillaries by an organosilane layer which has functional groups for anchoring peptidic or peptoid target molecules; c) defining locally delimited sample areas comprising a group of adjacent areas by selectively applying a reagent for protecting the functional groups or a protected anchor molecule for covalent bonding to the functional groups by a pipetting robot into the capillaries of the sample areas; d) chemically deactivating the regions of the capillary plate, which are outside the sample areas, with functional groups, by the application of a deactivation reagent; e) deprotecting all protected functional groups or anchor molecules; f) applying the amino-terminally and side chain-protected reagents required for production of the individual target molecules to the sample areas of the capillary plate selectively with the pipetting robot and covalently bonded to the de-protected functional groups or anchor molecules; g) removing excess reagents by way of a variable-volume cavity with a discharge at an underside of the synthesis table for disposal thereof, and h) repeating steps e) through g) are repeated as required.
 8. (amended) The method of claim 7, wherein γ-aminopropyltrialkoxysilane is used for silanisation in step b).
 9. (amended) The method of 8, wherein a protected α-aminopoly(ethyleneglycol)-ω-carboxylic acid active ester is used as the protected anchor molecule in step c).
 10. (amended) The method of claim 9, wherein fluorenylmethoxycarbonyl (FMOC)-protected α-aminopoly(ethyleneglycol)-ω-propionic acid-N-hydroxybenzotriazole ester is used as the protected anchor molecule in step c).
 11. (amended) A device for binding soluble or suspendable active substances to solid-phase bonded peptidic or peptoid target molecules, wherein the target molecules are bonded on substance libraries in the form of a planar structure with areas which are separated laterally from each other by physical barriers, the device comprising: an incubation table for fixing the structure in a defined position during binding of the active substances; and a closed variable volume cavity which is provided with a valve and into which liquids with the soluble or suspendable active substances and other liquids and gases can be introduced and removed, the variable volume cavity positioned on at least one side of the structure.
 12. (amended) The device of claim 11, wherein the structure comprises a capillary plate with a plurality of capillaries which serve to receive individual peptidic or peptoid target molecules of the substance library or the precursors thereof.
 13. (amended) The device of claim 12, further comprising: an ultrasonic transmitter which is in contact with the capillary plate.
 14. (amended) A device for investigating the interaction of soluble and suspendable active substances with solid-phase bonded peptidic or peptoid target molecules on substance libraries in the form of a planar structure which is freely accessible to optical measurement and having areas which are laterally separated from each other by physical barriers, the device comprising: an incubation table for fixing the structure in a defined position during the investigation in a defined position, and an optical measuring device having a radiation source for an excitation light and having a detection optical means.
 15. (amended) The device of claim 14, wherein the structure comprises a capillary plate with a plurality of capillaries which serve to receive individual peptidic or peptoid target molecules of the substance library or the precursors thereof.
 16. (amended) The device of claim 15, further comprising: an ultrasonic transmitter which is in contact with the capillary plate.
 17. (amended) The device of claim 16, wherein the optical measuring device comprises a fluorescence spectrometer.
 18. (amended) The device of claim 17, wherein the radiation source comprises a laser.
 19. (amended) A method of investigating the interaction of soluble and suspendable active substances with solid-phase bonded peptidic or peptoid target molecules on substance libraries in the form of a planar structure with areas which are laterally separated from each other by physical barriers, the method comprising the step of: providing an excitation light by way of an optical measuring device having a radiation source, which light scans the areas and is collected by way of a detection optical means.
 20. (amended) The method of claim 19, wherein the structure comprises a capillary plate having a plurality of capillaries which serve to receive individual peptidic or peptoid target molecules of the substance library or the precursors thereof.
 21. (amended) The method of claim 20, wherein the optical measuring device comprises a fluorescence spectrometer and the soluble or suspendable active substances and/or solid-phase bonded target molecules are fluorescence-marked.
 22. (amended) The method of claim 21 wherein the fluorescence marking is excited and detected with light in the UV, in the visible range or near infrared.
 23. (amended) The method of claim 22, wherein binding or non-binding of the active substance to the target molecule or the reaction of the active substance with the target molecule is detected by fluorescence or fluorescence extinction.
 24. (amended) The method of claim 23, wherein fluorescence excitation is effected with a pulsed diode laser with a pulse duration of less than 10 ns.
 25. (amended) The method of claim 23, wherein detection of the fluorescence is effected for the suppression of stray light in a time gate after decay of the excitation pulse.
 26. (new) The device of claim 1, wherein the structure has regions outside the sample areas with functional groups that are chemically deactivated.
 27. (new) The device of claim 4, wherein the planar structure is transparent or opaque and can be used for optical measurement.
 28. (new) The method of claim 7, wherein γ-aminopropyltrialkoxysilane is used for silanisation in step b).
 29. (new) The method of 8, wherein a protected α-aminopoly(ethyleneglycol)-ω-carboxylic acid active ester is used as the protected anchor molecule in step c).
 30. (new) The method of claim 9, wherein fluorenylmethoxycarbonyl (FMOC)-protected α-aminopoly(ethyleneglycol)-ω-propionic acid-N-hydroxybenzotriazole ester is used as the protected anchor molecule in step c).
 31. (new) The device of claim 14, wherein the optical measuring device comprises a fluorescence spectrometer.
 32. (new) The device of claim 17, wherein the radiation source comprises a laser.
 33. (new) The method of claim 20, wherein the optical measuring device comprises a fluorescence spectrometer and the soluble or suspendable active substances and/or solid-phase bonded target molecules are fluorescence-marked.
 34. (new) The method of claim 33 wherein the fluorescence marking is excited and detected with light in the UV, in the visible range or near infrared.
 35. (new) The method of claim 19, wherein binding or non-binding of the active substance to the target molecule or the reaction of the active substance with the target molecule is detected by fluorescence or fluorescence extinction.
 36. (new) The method of claim 34, wherein binding or non-binding of the active substance to the target molecule or the reaction of the active substance with the target molecule is detected by fluorescence or fluorescence extinction.
 37. (new) The method of claim 35, wherein fluorescence excitation is effected with a pulsed diode laser with a pulse duration of less than 10 ns.
 38. (new) The method of claim 36, wherein fluorescence excitation is effected with a pulsed diode laser with a pulse duration of less than 10 ns.
 39. (new) The method of claim 22, wherein fluorescence excitation is effected with a pulsed diode laser with a pulse duration of less than 10 ns.
 40. (new) The method of claim 34, wherein fluorescence excitation is effected with a pulsed diode laser with a pulse duration of less than 10 ns.
 41. (new) The method of claim 35, wherein detection of the fluorescence is effected for the suppression of stray light in a time gate after decay of the excitation pulse.
 42. (new) The method of claim 36, wherein detection of the fluorescence is effected for the suppression of stray light in a time gate after decay of the excitation pulse.
 43. (new) The method of claim 22, wherein detection of the fluorescence is effected for the suppression of stray light in a time gate after decay of the excitation pulse.
 44. (new) The method of claim 34, wherein detection of the fluorescence is effected for the suppression of stray light in a time gate after decay of the excitation pulse. 